Division of Biological Sciences, Neurobiology Section, University of California, San Diego, San Diego, United States; Howard Hughes Medical Institute, University of California, San Diego, San Diego, United States
Alexandr Goncharov
Division of Biological Sciences, Neurobiology Section, University of California, San Diego, San Diego, United States; Howard Hughes Medical Institute, University of California, San Diego, San Diego, United States
Mark H Ellisman
National Center for Research in Biological Systems, Department of Neurosciences, School of Medicine, University of California, San Diego, San Diego, United States
Division of Biological Sciences, Neurobiology Section, University of California, San Diego, San Diego, United States; Howard Hughes Medical Institute, University of California, San Diego, San Diego, United States
Subcellular localization of ribosomes defines the location and capacity for protein synthesis. Methods for in vivo visualizing ribosomes in multicellular organisms are desirable in mechanistic investigations of the cell biology of ribosome dynamics. Here, we developed an approach using split GFP for tissue-specific visualization of ribosomes in Caenorhabditis elegans. Labeled ribosomes are detected as fluorescent puncta in the axons and synaptic terminals of specific neuron types, correlating with ribosome distribution at the ultrastructural level. We found that axonal ribosomes change localization during neuronal development and after axonal injury. By examining mutants affecting axonal trafficking and performing a forward genetic screen, we showed that the microtubule cytoskeleton and the JIP3 protein UNC-16 exert distinct effects on localization of axonal and somatic ribosomes. Our data demonstrate the utility of tissue-specific visualization of ribosomes in vivo, and provide insight into the mechanisms of active regulation of ribosome localization in neurons.
